Effectiveness of different solid-phase microextraction fibres for differentiation of selected Madeira island fruits based on their volatile metabolite profile: identification of novel compounds

A headspace solid-phase microextraction (HS-SPME) procedure based on five commercialised fibres (85 μm polyacrylate – PA, 100 μm polydimethylsiloxane – PDMS, 65 μm polydimethylsiloxane/divinylbenzene – PDMS/DVB, 70 μm carbowax/divinylbenzene – CW/DVB and 85 μm carboxen/polydimethylsiloxane – CAR/PDMS) is presented for the characterization of the volatile metabolite profile of four selected Madeira island fruit species, lemon (Citrus limon), kiwi (Actinidia deliciosa), papaya (Carica papaya L.) and Chickasaw plum (Prunus angustifolia). The isolation of metabolites was followed by thermal desorption gas chromatography–quadrupole mass spectrometry (GC–qMS) methodology. The performance of the target fibres was evaluated and compared. The SPME fibre coated with CW/DVB afforded the highest extraction efficiency in kiwi and papaya pulps, while in lemon and plum the same was achieved with PMDS/DVB fibre. This procedure allowed for the identification of 80 compounds, 41 in kiwi, 24 in plums, 23 in papaya and 20 in lemon. Considering the best extraction conditions, the most abundant volatiles identified in kiwi were the intense aldehydes and ethyl esters such as (E)-2-hexenal and ethyl butyrate, while in Chicasaw plum predominate 2-hexenal, 2-methyl-4-pentenal, hexanal, (Z)-3-hexenol and cyclohexylene oxide. The major compounds identified in the papaya pulp were benzyl isothiocyanate, linalool oxide, furfural, hydroxypropanone, linalool and acetic acid. Finally, lemon was shown to be the most divergent of the four fruits, being its aroma profile composed almost exclusively by terpens, namely limonene, γ-terpinene, o-cymene and α-terpinolene. Thirty two volatiles were identified for the first time in the fruit or close related species analysed and 14 volatiles are reported as novel volatile metabolites in fruits. This includes 5 new compounds in kiwi (2-cyclohexene-1,4-dione, furyl hydroxymethyl ketone, 4-hydroxydihydro-2(3H)-furanone, 5-acetoxymethyl-2-furaldehyde and ethanedioic acid), 4 in plum (4-hydroxydihydro-2(3H)-furanone, 5-methyl-2-pyrazinylmethanol, cyclohexylene oxide and 1-methylcyclohexene), 4 in papaya (octaethyleneglycol, 1,2-cyclopentanedione, 3-methyl-1,2-cyclopentanedione and 2-furyl methyl ketone) and 2 in lemon (geranyl farnesate and safranal). It is noteworthy that among the 15 volatile metabolites identified in papaya, 3-methyl-1,2-cyclopentanedione was previously described as a novel PPARγ (peroxisome proliferator-activated receptor γ) agonist, having a potential to minimize inflammation.

EFEITO do ÁCIDO GIBERÉLICO (AG3) NA FLORAÇÃO E PRODUÇÃO da LIMA ÁCIDA 'TAHITI' (Citrus latifolia Tan.)

A floração nos citros, assim como em outras fruteiras, é um dos fatores determinantes para a produção. A possibilidade de inibição ou redução da florada normal e alteração da época de produção para a lima 'Tahiti' é fator primordial para determinar sua rentabilidade econômica. O presente trabalho teve como objetivo avaliar o efeito do ácido giberélico sobre a florada, número de flores formadas, da lima ácida 'Tahiti', nas condições do Estado de São Paulo durante dois anos, buscando inibir a florada normal e observar o efeito desta inibição sobre produções temporãs. O delineamento experimental foi em blocos casualizados (5 blocos) com 3 plantas por tratamento. Foi utilizada uma planta como bordadura entre os tratamentos e uma linha de plantas entre os blocos. Os tratamentos foram: testemunha, 20, 40 e 80 mg/L de ácido giberélico (AG3) + 10 ml/L de espalhante adesivo, e o pH da água utilizada foi 6,2. As plantas foram tratadas com ácido giberélico durante o inverno (estação seca) após 50 dias sem irrigação no primeiro ano e 60 dias no segundo. O ácido giberélico reduziu o número de flores formadas (-81%) e aumentou a produção de frutos temporões (+59,77% ou +16,04 kg/pl).

Desbaste da limeira-ácida-'Tahiti' (Citrus latifolia Tanaka) com TPA e efeitos na produção

Foram avaliadas concentrações de 0; 10; 20; 40 e 80 mg/L do 3,5,6-TPA (Ácid 3,5,6-tricloro-2-piridil- oxiacético), em sua formulação éster, para desbaste de frutos novos da limeira-ácida-'Tahiti' (Citrus latifolia Tanaka), nas condições ecológicas do Estado de São Paulo. O 3,5,6-TPA, na concentração 10mg/L, foi o tratamento mais eficaz para uso na cultura. Concentrações maiores causaram severos danos às plantas. Essa concentração, aplicada logo após a florada de outubro, reduziu em 65% a produção do mês de fevereiro seguinte. Todavia, as produções dos meses subseqüentes não diferiram entre si. A concentração de 10mg/L causou alongamento dos frutos, aumento da espessura da casca e redução do Brix, na época da colheita.

Susceptibility of citrus species to Alternaria alternata, the causal agent of the Alternaria brown spot

Alternaria alternata, the causal agent of Alternaria brown spot (ABS), causes necrosis on leaves, twigs, and fruit, reducing the productivity and quality of fruits. Tangerines and their hybrids are highly susceptible to the disease. Species, hybrids, and cultivars of Citrus from the germplasm bank of the Estacao Experimental de Citricultura de Bebedouro, São Paulo, Brazil, were evaluated in 2004 and 2005 with respect to their resistance to A. alternata, both through natural infection and by inoculation. Detached leaves were also used to demonstrate susceptibility or resistance to the disease. Ten cultivars of Satsumas (Citrus unshiu), and 14 cultivars of Clementine mandarin (C. clementina) did not show any symptoms of the disease in their leaves, either through natural infection or when inoculated in the field. The Burguess SRA-412, Wallent SRA-438, Carvalhais, Ampefy SRA-459, Ananas SRA, and Macaque SRA-426 mandarin hybrids (C. reticulata) did not show symptoms of the disease under natural or artificial infection in the field. Some cultivars of C. deliciosa, C. tangerina, C. erythrosa, and C. temple showed symptoms of the disease, even though no previous record of their susceptibility to Alternaria brown spot had been previously reported. The hybrids Fairchild, Nova, Page, Fortune, and Sunburst were susceptible to the disease. However, Fremont mandarin (a crossing between C. clementina and C. reticulata), Encore (C. nobilis x C. deliciosa), and Fallglo (C. reticulata x C. paradisi) did not show symptoms in field, and few symptoms were verified in detached leaves. These materials are promising for the cultivation of tangerines, and will enable genetic improvement for the development of cultivars resistant to Alternaria brown spot. (c) 2007 Elsevier B.V. All rights reserved.

Biofertilizer for control of Guignardia citricarpa, the causal agent of citrus black spot

Biofertilizers generated from the anaerobic and aerobic digestion of cattle manure, known as Bio1 and Bio2, respectively, were studied with regard to their microbial composition and effect on the mycelial growth of Phyllosticta citricarpa, the causal agent of citrus black spot (CBS). Two field experiments were conducted to determine the biofertilizer's potential (Bio1) in controlling CBS (2001/2002 and 2002/2003 crops). It was observed that the greatest number of microorganisms was found in the aerobically produced biofertilizer. Bio2 did not inhibit the mycelial growth of P. citricarpa. Mycelial growth of P. citricarpa was inversely proportional to the Bio1 biofertilizer concentration. In the 2001/2002 cropping season, the Biol effect in controlling CBS was directly proportional to its concentration, at the rate of 0 (healthy fruit), with R-2 = 0.88. Biol had a significant effect in controlling CBS, at a concentration of 10%, during the 2001/2002 cropping season, with DI values of 0.246 and 0.229 for the. 10 and 20% doses, respectively, compared to DI of 0.329 for the control. A directly proportional effect of the biofertilizer concentration on the percentage of fruits with a rating of zero was. also observed in the 2002/2003 cropping season, with R-2 = 0.48. However, even at doses higher than in the preceding cropping season, the biofertilizer was less effective, possibly due to a higher occurrence of the disease. Copper oxychloride and combined applications of copper oxychloride and carbendazim plus mancozeb controlled the disease. The possibility of using the biofertilizer as a protective biofungicide to replace copper oxychloride, especially in organic agriculture, should be explored. (c) 2005 Elsevier Ltd. All rights reserved.

The effects of inter-crop cultivation Between rows of citrus crop on spreading of Guignardia citricarpa Ascospores and in the citrus black spot occurrence

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Susceptibility of fruits of the 'Valência' and 'Natal' sweet orange varieties to Guignardia citricarpa and the influence of the coexistence of healthy and symptomatic fruits

Em pomar de laranjeiras 'Valência' e 'Natal' avaliou-se a importância da presença de frutos sintomáticos da mancha preta citros (MPC) na severidade da doença nos frutos cítricos da safra subseqüente. Adicionalmente, avaliou-se o estádio de suscetibilidade dos frutos dessas variedades. Frutos foram protegidos com sacos de papel cristal a partir do estádio de 75% de pétalas caídas em outubro de 2000, até abril de 2001. Frutos foram expostos, em intervalos semanais, da 1ª à 24ª semana. Esse processo se deu tanto em plantas onde os frutos da safra remanescente foram previamente colhidos, como naquelas cujos frutos sintomáticos da safra remanescente permaneceram até a sua queda natural. Avaliou-se a severidade da doença usando uma escala de notas que variou de 0 (ausência de sintomas) a 6 (sintomas severos). Observou-se que para as duas variedades os conídios de Phyllosticta citricarpa, formados nas lesões dos frutos da safra remanescente, não provocaram incremento significativo na severidade da doença dos frutos da safra subseqüente. A proteção dos frutos até 10ª semana após a queda de pétalas não influenciou na quantidade final de lesões, indicando que as descargas de ascósporos que ocorreram a partir desse momento foram, provavelmente, responsáveis pela severidade da doença. Frutos que ficaram expostos entre a 20ª a 24ª semanas após a queda de 75% de pétalas mostraram-se sintomáticos, indicando que nesse estádio frutos encontravam-se suscetíveis ao patógeno.

Postharvest harpin or Bacillus thuringiensis treatments suppress citrus black spot in 'Valencia' oranges

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Acaricides used in conventional and organic citrus orchard: leprosis and phytoseiid mites population control

The citrus leprosis control in São Paulo state is performed exclusively by acaricides to control the vector mite, Brevipalpus phoenicis, which increases the production costs and may affect the beneficial organism's population. Therefore, the aim of this trial was to evaluate during four seasons, the effects of acaricides recommended to control the mite B. phoenicis in conventional and organic citrus over evolution of citrus leprosis and over phytoseiids' population. The experiment was installed in October of 2003 in a citrus orchard in Reginopolis city, state of São Paulo. The experimental design used randomized blocks, the dosages was expressed as mL c.p./100L of water and the treatments were the following: spirodiclofen (20 mL); cyhexatin (50 mL) (used in rotation), lime sulfur (4,000 mL) and control (without pesticide application). However, the rotation between spirodiclofen and cyhexatin began in September 2006. Prior to that time, only spirodiclofen had been used. Surveys were conducted every 15 days on the B. phoenicis, Iphiseioides zuluagai, and Euseius populations. The control level adopted by the B. phoenicis was 8.3%, and the pesticide applications were conducted using tractor-sprayers. During the 2007-08 seasons, 10 infected fallen fruits per plot were collected and the number of leprosis lesions was quantified by each fruit. By the end of the 2007-08 seasons, the productivity, harvest losses, the disease incidence and severity were evaluated. It was found that the lesions' location over the fruit is more important in determining its drop than the lesions' number. The more intense the mite infestation, the greater is the number of lesions, resulting in increased premature fruit drop. A strategy using acaricides spirodiclofen and cyhexatin in rotation promoted more efficient control of B. phoenicis compared to lime sulfur, resulting in greater productivity, lower fruit losses and severity levels. The lime sulfur applications reduced the mite population incidence below the control level; however it did not prevent the lesions' occurrence. The acaricides applicarevented adverse effects on phytoseiid population because there was a reduction of their density.

A new method for inoculation of fruit with Guignardia citricarpa, the causal agent of citrus black spot

Citrus black spot (CBS) is a fungal disease, caused by Guignardia citricarpa, that has a high economic impact on citrus. Although G. citricarpa has been associated with black spot of citrus, an adequate pathogenicity test is still not available. Thus, our objective was to develop and evaluate a simple, safe, and practical pathogenicity test. We used fruits from Pera-Rio and Valencia sweet orange trees from two different orchards, located in the State of São Paulo, Brazil. Inoculation was performed by placing six disks colonized by G. citricarpa, onto the peel of healthy fruits, previously bagged. In the Pera-Rio sweet orange grove, initial symptoms of the false melanose type resulting from the inoculations were observed 55 days after inoculation (dai). In the Valencia grove, initial symptoms also of the false melanose type resulting from the inoculations occurred 73 dai. A total of 92.8% and 86.6% of the Pera Rio and Valencia fruits inoculated, respectively, showed symptoms of CBS. Citrus black spot symptoms were not observed in any of the control fruits.

A simple and non-destructive model for individual leaf area estimation in citrus

Introduction. Leaf area is often related to plant growth, development, physiology and yield. Many non-destructive models have been proposed for leaf area estimation of several plant genotypes, demonstrating that leaf length, leaf width and leaf area are closely correlated. Thus, the objective of our study was to develop a reliable model for leaf area estimation from linear measurements of leaf dimensions for citrus genotypes. Materials and methods. Leaves of citrus genotypes were harvested, and their dimensions (length, width and area) were measured. Values of leaf area were regressed against length, width, the square of length, the square of width and the product (length x width). The most accurate equations, either linear or second-order polynomial, were regressed again with a new data set; then the most reliable equation was defined. Results and discussion. The first analysis showed that the variables length, width and the square of length gave better results in second-order polynomial equations, while the linear equations were more suitable and accurate when the width and the product (length x width) were used. When these equations were regressed with the new data set, the coefficient of determination (R(2)) and the agreement index 'd' were higher for the one that used the variable product (length x width), while the Mean Absolute Percentage Error was lower. Conclusion. The product of the simple leaf dimensions (length x width) can provide a reliable and simple non-destructive model for leaf area estimation across citrus genotypes.

Fertirrigação com diferentes doses de NPK e seus efeitos sobre a produção e qualidade de frutos de laranja (Citrus sinensis O.) 'Valência'

O experimento foi realizado na Fazenda Três Irmãos, no município de Pratânia, Estado de São Paulo, com o objetivo de estudar o efeito da fertirrigação com diferentes doses de NPK aplicadas por meio de um sistema de irrigação por microaspersão na cultura da laranja em comparação com a adubação convencional irrigada e não irrigada. A avaliação dos efeitos da irrigação e da fertirrigação na cultura foi feita com base na produção e qualidade dos frutos colhidos, na análise de solo da camada de 0-20 cm e na análise de folhas. Os tratamentos não influenciaram significativamente a produtividade de acordo com a análise estatística realizada, embora se tenha observado uma tendência de maior produtividade nos tratamentos de fertirrigação para uma mesma dose. A análise de qualidade dos frutos não mostrou diferenças significativas entre os tratamentos. Os teores de alguns dos elementos obtidos na análise de solo apresentaram diferenças significativas proporcionadas pela irrigação e/ou modo de aplicação dos fertilizantes à cultura. A análise de folhas diferiu quanto aos teores de Ca, Mg e K.

Gastric Ulcers in Middle-Aged Rats: The Healing Effect of Essential Oil from Citrus aurantium L. (Rutaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Phytochemicals and Antioxidant Activity of Citrus Seed Oils

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Modelagem matemática aplicada ao controle biológico de Ceratitis capitata Wiedemann (Diptera: Tephritidae) por Diachasmimorpha longicaudata Ashmed (Hymenoptera: Braconidae) em Citrus

O modelo matemático apresentado tem como objetivos: (1) simular as dinâmicas populacionais de um sistema hospedeiro parasitóide de três níveis tróficos composto pelas populações de mosca-do-mediterrâneo Ceratitis capitata (Wiedemann), vespa braconídea parasitóide Diachasmimorpha longicaudata (Ashmed) e frutos cítricos; (2) auxiliar no melhor entendimento dos principais fatores biológicos e ecológicos que regem as interações populacionais e (3) colaborar com programas mais eficientes de controle biológico para o sistema em questão. A metodologia empregada baseou-se na formulação de sistemas de equações de diferenças que descrevessem os processos de interação do sistema trófico. Posteriormente, foram elaboradas resoluções numéricas desses sistemas de equações e sua representação gráfica, utilizando-se o programa computacional Matlab, versão 6.1. Os dados biológicos e ecológicos necessários para a formulação das equações matemáticas foram fornecidos por especialistas em controle de C. capitata e retirados da literatura referente ao controle biológico das moscas-das-frutas em plantações de citros no Brasil, principalmente através da utilização de vespas parasitóides, como D. longicaudata. Os resultados obtidos nas simulações sugerem que o modelo proposto descreve adequadamente o sistema ecológico em questão e permite entender melhor suas principais características biológicas e ecológicas. em conseqüência pode auxiliar na escolha do modo e momento para liberação da vespa parasitóide para o controle mais efetivo de C. capitata.